Assembly of an abundant endogenous major histocompatibility complex class II/peptide complex in class II compartments

To identify the intracellular site(s) of formation of an endogenous class II/peptide complex in a human B cell line, we employed kinetic pulse-chase labeling experiments followed by subcellular fractionation by Percoll density gradient centrifugation and immunogold labeling on ultrathin cryosections. For direct demonstration of assembly of such complexes, we used the monoclonal antibody YAe, which detects an endogenous complex of the mouse class II molecule I-Ab with a 17-amino acid peptide derived from the alpha chain of HLA-DR (DR alpha52-68). We show that in human B lymphocytes, these class II/peptide complexes assemble and transiently accumulate in major histocompatibility complex class II-enriched compartments before reaching the cell surface.     

Gilthead Seabream Liver Integrative Proteomics and Metabolomics Analysis Reveals Regulation by Different Prosurvival Pathways in the Metabolic Adaptation to Stress

The study of the molecular mechanisms of stress appraisal on farmed fish is paramount to ensuring a sustainable aquaculture. Stress exposure can either culminate in the organism’s adaptation or aggravate into a metabolic shutdown, characterized by irreversible cellular damage and deleterious effects on fish performance, welfare, and survival. Multiomics can improve our understanding of the complex stressed phenotype in fish and the molecular mediators that regulate the underlying processes of the molecular stress response. We profiled the stress proteome and metabolome of Sparus aurata responding to different challenges common to aquaculture production, characterizing the disturbed pathways in the fish liver, i.e., the central organ in mounting the stress response. Label-free shotgun proteomics and untargeted metabolomics analyses identified 1738 proteins and 120 metabolites, separately. Mass spectrometry data have been made fully accessible via ProteomeXchange, with the identifier PXD036392, and via MetaboLights, with the identifier MTBLS5940. Integrative multivariate statistical analysis, performed with data integration analysis for biomarker discovery using latent components (DIABLO), depicted the 10 most-relevant features. Functional analysis of these selected features revealed an intricate network of regulatory components, modulating different signaling pathways related to cellular stress, e.g., the mTORC1 pathway, the unfolded protein response, endocytosis, and autophagy to different extents according to the stress nature. These results shed light on the dynamics and extent of this species’ metabolic reprogramming under chronic stress, supporting future studies on stress markers’ discovery and fish welfare research.     

Influence of Different Cooking Methods on the Concentration of Glucosinolates and Vitamin C in Broccoli

Broccoli belongs to the Brassicaceae family and the Brassica genus, also designated crucifers, which has been linked to reduced risk of certain diseases for their content of compounds like glucosinolates (GLS) and vitamin C. Isothiocyanates, nitriles, and thiocyanates are degradation products of glucosinolates, are substances that protect cells against oxidative stress, and present many other health benefits. Vitamin C also contains antioxidant properties that contribute to the beneficial effects that broccoli have for health. The present paper is an overview of the reduction of secondary plant products, such as GLS and vitamin C, by cooking. Nonetheless, these concentrations can be modified if the cell structure of the plant is disrupted, e.g., while cutting, chewing, or cooking. Myrosinase can come into contact with GLS and hydrolyze it to isothiocyanate sulforaphane or sulforaphane nitrile, depending on the environmental conditions, which produces changes in the composition and concentration of GLS. Thus, cooking induces many chemical and physical modifications in food, among which GLS and vitamin C content in broccoli can change. Vitamin C and GLS are water-soluble, which makes them more susceptible to loss during the cooking process. Despite some controversy, most reviewed studies show that conventional cooking methods (boiling, steaming, and frying) and nonconventional ones (microwaving) significantly lead to the degradation of vitamin C and GLS. Nonetheless, steaming is the conventional method that obtains the best result to better preserve these two compounds.     

Performance evaluation of an anaerobic fluidized bed reactor with natural zeolite as support material when treating high-strength distillery wastewater

The performance of two laboratory-scale fluidized bed reactors with natural zeolite as support material when treating high-strength distillery wastewater was assessed. Two sets of experiments were carried out. In the first experimental set, the influences of the organic loading rate (OLR), the fluidization level (FL) and the particle diameter of the natural zeolite (D_P) were evaluated. This experimental set was carried out at an OLR from 2 to 5 g COD (chemical oxygen demand)/l d, at FL 20% and 40% and with D_P in the range of 0.2–0.5 mm (reactor 1) and of 0.5–0.8 mm (reactor 2). It was demonstrated that OLR and FL had a slight influence on COD removal, whereas they had a strong influence on the methane production rate. The COD removal was slightly higher for the highest particle diameter used. The second experimental set was carried out at an OLR from 3 to 20 g COD/l d with 25% of fluidization and D_P in the above-mentioned ranges for reactors 1 and 2. The performance of the two reactors was similar; no significant differences were found. The COD removal efficiency correlated with the OLR based on a straight line. COD removal efficiencies higher than 80% were achieved in both reactors without significant differences. In addition, a straight line equation with a slope of 1.74 d⁻¹ and an intercept on the y-axis equal to zero described satisfactorily the effect of the influent COD on the COD removal rate. It was also observed that both COD removal rate and methane production (Q_M) increased linearly with the OLR, independently of the D_P used.     

Synthesis and optical properties of novel, thermally stable phenanthrolines bearing an arylthienyl-imidazo conjugation pathway

Six new hyperpolarizable chromophores based on functionalized arylthiophene donors and an imidazo-phenanthroline acceptor moiety have been designed and synthesized for the first time in good to excellent yields by condensation of 5,6-phenantroline-dione with formyl-arylthiophene derivatives in the presence of ammonium acetate in glacial acetic acid. The thermal stability, solvatochromic and nonlinear optical properties of these compounds were evaluated.     

A novel tetanus neurotoxin-insensitive vesicle-associated membrane protein in SNARE complexes of the apical plasma membrane of epithelial cells

The importance of soluble N-ethyl maleimide (NEM)-sensitive fusion protein (NSF) attachment protein (SNAP) receptors (SNAREs) in synaptic vesicle exocytosis is well established because it has been demonstrated that clostridial neurotoxins (NTs) proteolyze the vesicle SNAREs (v-SNAREs) vesicle-associated membrane protein (VAMP)/brevins and their partners, the target SNAREs (t-SNAREs) syntaxin 1 and SNAP25. Yet, several exocytotic events, including apical exocytosis in epithelial cells, are insensitive to numerous clostridial NTs, suggesting the presence of SNARE-independent mechanisms of exocytosis. In this study we found that syntaxin 3, SNAP23, and a newly identified VAMP/brevin, tetanus neurotoxin (TeNT)-insensitive VAMP (TI-VAMP), are insensitive to clostridial NTs. In epithelial cells, TI-VAMP-containing vesicles were concentrated in the apical domain, and the protein was detected at the apical plasma membrane by immunogold labeling on ultrathin cryosections. Syntaxin 3 and SNAP23 were codistributed at the apical plasma membrane where they formed NEM-dependent SNARE complexes with TI-VAMP and cellubrevin. We suggest that TI-VAMP, SNAP23, and syntaxin 3 can participate in exocytotic processes at the apical plasma membrane of epithelial cells and, more generally, domain-specific exocytosis in clostridial NT-resistant pathways.     

The synthesis and characterization of heterocyclic azo dyes derived from 5-N,N-dialkylamino-2,2′-bithiophene couplers

Heterocyclic azo dyes were synthesized by diazotation of several substituted anilines and coupling with 5-N,N-dialkylamino-2,2′-bithiophenes to give 4-phenylazo-5-N,N-dialkylamino-2,2′-bithiophenes. This reaction contrasts with the behavior of 5-alkoxy-2,2′-bithiophenes towards aryldiazonium salts which provides 5-phenylazo-5-alkoxy-2,2′-bithiophenes. The thermal stability of the derivatives was evaluated using thermogravimetric analysis and their solvatochromic behaviour was investigated in several solvents of different polarity. The experimental results indicate that the heterocyclic azo dyes could be used as thermally stable, solvatochromic probes.     

Performance and kinetic evaluation of the anaerobic digestion of two-phase olive mill effluents in reactors with suspended and immobilized biomass

A lab-scale study was conducted on the mesophilic anaerobic digestion of two-phase olive mill effluents constituted by the mixture of the wash waters derived from the initial cleansing of the olives and those obtained in the washing and purification of virgin olive oil. The digestion was conducted in two continuously stirred tank reactors, one with biomass immobilized on Bentonite (reactor B) and other with suspended biomass used as control (reactor C). The reactors B and C operated satisfactorily between hydraulic retention times of 25.0 and 4.0 days and 25.0 and 5.0 days, respectively. Total chemical oxygen demand (TCOD) efficiencies in the ranges of 88.8-72.1% and 87.9-71.2% were achieved in the reactors with immobilized and suspended biomass, respectively, at organic loading rates of between 0.86 and 5.38 g TCOD/ld and 0.86-4.30 g TCOD/ld, respectively. On comparing both reactors for the same OLRs applied, it was observed that the reactor with support was always more efficient and stable showing higher TCOD, SCOD removal efficiencies and lower VFA/alkalinity ratio values than those found in the control reactor. A mass (TCOD) balance around the reactors allowed the methane yield coefficient, Y(G/S), to be obtained, which gave values of 0.31 and 0.30l CH(4)/g TCOD(removed) for reactors B and C, respectively. The cell maintenance coefficients,k(m), obtained by means of this balance were found to be 0.0024 and 0.0036 g TCOD(removed)/g VSSd, respectively. The volumetric methane production rates correlated with the biodegradable TCOD concentration through an equation of the Michaelis-Menten type for the two reactors studied. This proposed model predicted the behavior of the reactors very accurately showing deviations lower than 10% between the experimental and theoretical values of methane production rates.